Project Summary DNASU is non-profit repository that provides academia, the government and industry with DNA-based resources, which comprise a wide variety of plasmids including specialized experimental vectors, regulatory genetic sequences, and genes from over 1,100 organisms. The plasmids stored at DNASU required significant time and cost to produce, and DNASU seeks to avoid duplicative effort and to accelerate research by sharing them openly. In addition, DNASU safely archives the plasmids and gathers detailed annotation for each in a uniform and easily accessible format. Users access the repository through our website, https://dnasu.org/, where they can view pertinent clone, gene and vector information, as well as link-outs to external resources such as protocols and publications. DNASU acquires plasmids both through in-house production by our own high throughput cloning pipeline and through deposition by outside research groups, including the ORFeome Collaboration, which is a multinational collaboration that seeks to build a complete set of human genes and share them. In addition, we partnered with the Protein Structure Initiative (PSI), for whom we performed full- length sequencing and annotation on plasmids from PSI centers before depositing them. All plasmids are stored in both an automated storage retrieval system (ASRS) as a working collection and in separately located - 80? C freezers for archived storage. Distribution is initiated when a user visits DNASU to find and request selected construct(s) via the Internet. The electronic request is passed to the ASRS, which retrieves the clones for a technician who subcultures and ships the glycerol stocks for a nominal re-charge fee. DNASU maintains over 219,000 unique plasmids, and is constantly growing. The DNASU website sees an average of 9,000 hits, receives 155 orders and distributes over 650 plasmids per month. We have shipped >300,000 samples to scientists in 50 countries, with ~80% of these to North America. DNASU received funding from NIGMS through the PSI initiative that enabled the addition of >92,000 plasmids to its collection and the development of software tools for locating and using clones, thus creating a unique worldwide resource. For the next steps, we will strive to achieve self-sustainability utilizing the following four specific aims that address the operation and maintenance facets of DNASU: 1) Update and modernize DNASU software to current specifications; 2) Complete the Human ORFeome collection to establish the first complete human gene set collection; 3) Enhance our plasmid maintenance, archives and standard operating procedures to ensure the highest quality control; and 4) Establish a marketing program to maintain community connectivity and increase awareness of DNASU as a routine tool for scientists. We will use a combination of adjustments to our re-charge fees and increased distribution to meet our financial goals and establish an advisory board to provide useful feedback on our progress. We seek funding from this PAR to assist in this plan to establish self-sufficiency so that DNASU can continue as a resource for many years to come.